Luna® Direct Probe One-Step RT-qPCR 4X Mix with UDG (No ROX)
The Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG (No ROX) enables robust RT-qPCR performance from challenging sample types and crude lysates. Enhanced inhibitor tolerance and a concentrated 4X formulation support higher sample input for improved assay sensitivity. Enhanced inhibitor tolerance and a concentrated 4X formulation enable higher sample input for improved assay sensitivity.
For instruments that do not require the ROX passive reference dye
Enhanced inhibitor tolerance delivers reliable amplification from both crude and purified samples
Accelerate qPCR workflows with extension times as short as 10 seconds
Increase assay sensitivity with a 4X formulation that supports higher sample input
Maximize your throughput by multiplexing up to 6 targets
Luna WarmStart® RT paired with Hot Start Taq increases reaction specificity and robustness, enabling room temperature setup
Reduce the risk of carryover contamination with thermolabile UDG and dUTP included in the mix
Eliminate pipetting errors with non-interfering, visible blue tracking dye
The Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG (No ROX) enables robust RT-qPCR performance from challenging sample types and crude lysates, combining enhanced inhibitor tolerance with a concentrated 4X formulation that supports higher sample inputs for improved assay sensitivity. It supports multiplex detection of up to 6 targets in a single reaction. The mix includes Luna WarmStart Reverse Transcriptase, Hot Start Taq DNA Polymerase, dNTPs, and Murine RNase Inhibitor in an optimized buffer. Combining Hot Start Taq with a novel, WarmStart-activated RT minimizes non-specific amplification, improves reaction specificity, and allows for room temperature setup. The engineered Luna WarmStart RT offers high thermostability, supporting an optimal reaction temperature of 55°C.
Thermolabile UDG and dUTP help prevent carryover contamination. Thermolabile UDG is inactivated above 50°C, eliminating any impact on qPCR performance.
This formulation contains no ROX reference dye and is compatible with any instrument that does not require ROX.
Luna One Step RT-qPCR (No ROX) product comparison
Figure 1: Multiplex detection (6 targets) with the Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG
Multiplex RT-qPCR was performed using the Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG over a 5-log range of Jurkat total RNA (100 ng to 10 pg) on a qTOWERiris® 96 UV ready real-time instrument. Amplification standard curves and efficiencies for each of the six human targets are shown. Reactions (20 μl) included primers and probes at 200 nM each and followed the product-recommended cycling conditions. All six targets were detected linearly in multiplex format, demonstrating strong amplification efficiency and high R² values.
Figure 2: Reliable amplification from multiple purified and crude sample types
RT-qPCR targeting Heme from human blood, PMMoV from wastewater, CymMV from Dendrobium Spanish Eyes was performed using the Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG. Performance with purified and crude samples in 20 μl reactions was evaluated:
Purified samples: 60 ng–60 pg RNA from K2-EDTA Human blood; 300 ng–30 pg RNA from wastewater; 100 ng–10 pg RNA from Dendrobium Spanish Eyes with a known CymMV infection.
Direct samples: 10, 15 and 20% K2-EDTA Human blood; 5, 10 and 20% composite wastewater, 1 mM leaf punch.
Sensitive, robust performance can be observed across a 5-log dilution of purified nucleic acid from various sample types. In addition, Luna Direct robustly amplifies targets directly from crude sample types. In all cases, NTC amplification was not observed.
Figure 3: Luna Direct enables robust cross-platform performance of a multiplex infectious disease assay
Multiplex RT-qPCR targeting Zika virus (POLY gene), Dengue virus serotype 3 (E gene), and human RNase P (RPPH1 gene) was performed using Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG in 20 μl reactions. Assay performance was evaluated across various real-time PCR instruments using a 5-log dilution series (10–100,000 copies per reaction) of Quantitative Synthetic RNA from Zika virus (ATCC VR-3252SD) and Dengue virus type 3 (ATCC VR-3230SD) diluted in 5 ng Jurkat total RNA (BioChain, #R1255815-50). Consistent, sensitive amplification was observed for each target across all platforms, demonstrating robust multiplex performance over a broad dynamic range.
A non-fluorescent visible blue tracking dye is included to assist in pipetting into clear vessels. The tracking dye does not overlap spectrally with fluorophores commonly used in qPCR and does not interfere with real-time detection.
The Luna Direct Probe One-Step RT-qPCR 4X Mix with UDG (No ROX) contains an inert visible blue tracking dye to eliminate pipetting errors
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